Development of a Golden Gate Assembly-Based Genetic Toolbox for Lactiplantibacillus plantarum and Its Application for Engineering Monoterpenoid Biosynthesis.

Lactiplantibacillus plantarum is a food-grade lactic acid bacterium widely used in the food and beverage industry. Recently, this probiotic organism has been applied as a biofactory for the production of pharmaceutical and food-related compounds, but existing promoters and expression vectors for the genetic engineering of L. plantarum rely on inefficient cloning strategies and are usually not well-characterized. We therefore developed a modular and standardized Golden Gate Assembly-based toolbox for the de novo assembly of shuttle vectors from Escherichia coli to L. plantarum. A collection of the most relevant genetic parts, e.g., different origins of replication and promoters, was incorporated in our toolbox and thoroughly characterized by flow cytometry and the fluorescence assay. Standardized fusion sites allow combining the genetic part freely into a plasmid in one step. This approach allows for the high-throughput assembly of numerous constructs in a standardized genetic context, thus improving the efficiency and predictability of metabolic engineering in L. plantarum. Using our toolbox, we were able to produce the aroma compounds linalool and geraniol in L. plantarum by extending its native mevalonate pathway with plant-derived monoterpenoid synthases.

Heterofermentative Lentilactobacillus buchneri and low dry matter reduce high-risk antibiotic resistance genes in corn silage by regulating pathogens and mobile genetic element.

The study of antibiotic resistance in the silage microbiome has attracted initial attention. However, the influences of lactic acid bacteria inoculants and dry matter (DM) content on antibiotic resistance genes (ARGs) reduction in whole-plant corn silage remain poorly studied. This study accessed the ARGs' risk and transmission mechanism in whole-plant corn silage with different DM levels and treated with Lactiplantibacillus plantarum or Lentilactobacillus buchneri. The macrolide and tetracycline were the main ARGs in corn silage. The dominant species (Lent. buchneri and Lactobacillus acetotolerans) were the main ARGs carriers in whole-plant corn silage. The application of Lent. buchneri increased total ARGs abundance regardless of corn DM. Whole-plant corn silage with 30 % DM reduced the abundances of integrase and plasmid compared with 40 % DM. The correlation and structural equation model analysis demonstrated that bacterial community succession, resulting from changes in DM content, was the primary driving factor influencing the ARGs distribution in whole-plant corn silage. Interestingly, whole-plant corn silage inoculated with Lent. buchneri reduced abundances of high-risk ARGs (mdtG, mepA, tetM, mecA, vatE and tetW) by regulating pathogens (Escherichia coli), mobile genetic elements (MGEs) genes (IS3 and IS1182), and this effect was more pronounced at 30 % DM level. In summary, although whole-plant corn silage inoculated with Lent. buchneri increased the total ARGs abundance at both DM levels, it decreased the abundance of high-risk ARGs by reducing the abundances of the pathogens and MGEs, and this effect was more noticeable at 30 % DM level.

Optimizing the fermentation parameters in the Lactic Acid Fermentation of Legume-based Beverages- a statistically based fermentation.

BACKGROUND: The market for beverages is highly changing within the last years. Increasing consumer awareness towards healthier drinks led to the revival of traditional and the creation of innovative beverages. Various protein-rich legumes were used for milk analogues, which might be also valuable raw materials for refreshing, protein-rich beverages. However, no such applications have been marketed so far, which might be due to unpleasant organoleptic impressions like the legume-typical "beany" aroma. Lactic acid fermentation has already been proven to be a remedy to overcome this hindrance in consumer acceptance. RESULTS: In this study, a statistically based approach was used to elucidate the impact of the fermentation parameters temperature, inoculum cell concentration, and methionine addition on the fermentation of lupine- and faba bean-based substrates. A total of 39 models were found and verified. The majority of these models indicate a strong impact of the temperature on the reduction of aldehydes connected to the "beany" impression (e.g., hexanal) and on the production of pleasantly perceived aroma compounds (e.g., ß-damascenone). Positively, the addition of methionine had only minor impacts on the negatively associated sulfuric compounds methional, dimethyl sulfide, dimethyl disulfide, and dimethyl trisulfide. Moreover, in further fermentations, the time was added as an additional parameter. It was shown that the strains grew well, strongly acidified the both substrates (pH ≤ 4.0) within 6.5 h, and reached cell counts of > 9 log10 CFU/mL after 24 h. Notably, most of the aldehydes (like hexanal) were reduced within the first 6-7 h, whereas pleasant compounds like ß-damascenone reached high concentrations especially in the later fermentation (approx. 24-48 h). CONCLUSIONS: Out of the fermentation parameters temperature, inoculum cell concentration, and methionine addition, the temperature had the highest influence on the observed aroma and taste active compounds. As the addition of methionine to compensate for the legume-typical deficit did not lead to an adverse effect, fortifying legume-based substrates with methionine should be considered to improve the bioavailability of the legume protein. Aldehydes, which are associated with the "beany" aroma impression, can be removed efficiently in fermentation. However, terminating the process prematurely would lead to an incomplete production of pleasant aroma compounds.

Unveiling the antimicrobial and antibiofilm potential of biosurfactant produced by newly isolated Lactiplantibacillus plantarum strain 1625.

The present study aimed to characterize the biosurfactants synthesized by lactic acid bacteria (LAB) obtained from fermented foods, optimize the conditions for increasing the yield of biosurfactants and explore their antimicrobial and antibiofilm potential. Out of the 26 LAB isolates, isolate BS2 showed the highest biosurfactant production as indicated in the oil displacement test, drop collapse and emulsification activity. BS2 was identified as Lactiplantibacillus plantarum 1625 using 16S-rRNA gene sequencing and phylogenetic analysis. The biosurfactant produced by BS2 was identified as an anionic glycol-lipo-proteins by employing Fourier Transform Infrared Spectroscopy (FTIR) and Gas Chromatography-Mass Spectrometry (GC-MS) analysis. The biosurfactants produced by L. plantarum 1625 demonstrated strong antibacterial and antibiofilm characteristics against pathogenic strains such as Staphylococcus aureus MTCC 1049, Escherichia coli MTCC 1587, and Pseudomonas putida MTCC 1655. The minimal inhibition concentration value of antibacterial activity was found to be 0.1 mg/mL with the inhibition percentage ranging from 90 to 95%. Further, the effect of temperature, pH, and substrate composition on biosurfactant production was also studied to enhance it production using the Box-Behnken Design approach of Response surface methodology (RSM). Application of biosurfactant led to a considerable decrease in biofilm-forming harmful bacteria, as proven by scanning electron microscopy analysis. The results highlight the potential uses of biosurfactants in distinct industries, and biotechnological contexts, especially in the creation of new antimicrobial and antibiofilm agents.

Responses of microbial community composition and CAZymes encoding gene enrichment in ensiled Elymus nutans to altitudinal gradients in alpine region.

High-throughput metagenomic sequence technology was employed to evaluate changes in microbial community composition and carbohydrate-active enzymes encoding gene enrichment status in Elymus nutans silages to altitudinal gradients in the world's highest alpine region of Qinghai-Tibetan Plateau (QTP). E. nutans were collected from three different altitudes in QTP: 2,600 m (low altitude), 3600 m (moderate altitude), and 4,600 m [high (H) altitude], and ensiled for 7, 14, 30, and 60 d. Results indicated an improvement in silage quality with the increasing altitude, although the acetic acid concentration and dry matter loss were greater in H altitude silages after 30 d of ensiling. Harmful bacteria or potential pathogens predominated in the microbial community on d 7 and 14 of fermentation, while genera belonging to lactic acid bacteria gradually became the main microorganisms with the increasing altitude on d 30 and 60 of ensiling. The abundance of carbohydrate-active enzymes genes responsible for macromolecular carbohydrate degradation in silage increased with increasing altitude, and those genes were mainly carried by Lactiplantibacillus and Pediococcus at 30 and 60 d of ensiling. The abundance of key enzymatic genes associated with glycolysis and organic acid production in carbohydrate metabolism pathway was higher in H altitude silages, and Lactiplantibacillus and Pediococcus were also the main hosts after 30 d of silage fermentation, except for the fact that acetic acid production was also related to genera Leuconostoc, Latilactobacillus, and Levilactobacillus. IMPORTANCE: The fermentation quality of Elymus nutans silage was getting better with the increase of altitude in the Qinghai-Tibetan Plateau. The abundance of hosts carrying carbohydrate-active enzymes genes and key enzyme genes related to organic acid production increased with increasing altitude during the later stages of fermentation. Lactiplantibacillus and Pediococcus were the core microorganisms responsible for both polysaccharide hydrolysis and silage fermentation in the late stage of ensiling. This study provided insights on the influence of different altitudes on the composition and function of silage microbiome in the Qinghai-Tibetan Plateau, and provided a reference approach for improving the quality and controllability of silage production in high altitude areas of the Qinghai-Tibetan Plateau.

Lactic acid bacteria isolated from women' breast milk and infants' faeces have appreciable immunogenic and probiotic potentials against diarrheagenic E. coli strains.

Diarrheal diseases remain the leading cause of high mortality among the infants, particularly in the developing countries; Probiotic intervention for diarrhea has been an ongoing novel approach to diarrheal prevention and treatment. This study aims to characterize immunogenic and probiotic properties of lactic acid bacteria (LAB) isolated from human breast milk and neonates' faeces. The LAB isolates from 16 mothers' breast milk and 13 infants' faeces were screened and identified by 16 S rRNA gene partial sequencing. Their antimicrobial activities against 5 strains of diarrheagenic Escherichia coli were tested. Organic acids production was quantified by HPLC, and antibiotic resistance pattern were determined by VITEK®. Autoaggregation, co-aggregation and hydrophobicity properties were assessed by UV spectrophotometry and immunomodulatory effect was determined in mouse model. Ninety-three LAB of five genera were identified. The most abundant species was Lactiplantibacillus plantarum with inhibition zones ranged from 8.0 to 25.0 ± 1 mm. Lacticaseibacillus rhamnosus A012 had 76.8 mg/mL lactic acid, (the highest concentration), was susceptible to all antibiotics tested. L. plantarum A011 and L. rhamnosus A012 were highly resistance to gastrointestinal conditions. L. rhamnosus A012 produced hydrophobicity of 25.01% (n-hexadecane), 15.4% (xylene) and its autoaggregation was 32.52%. L. rhamnosus A012 and L. plantarum A011 exert immunomodulatory effects on the cyclophosphamide-treated mice by upregulating anti-inflammatory cytokine and downregulating proinflammatory cytokines. Lactobacillus sp. demonstrated good probiotic and immunomodulatory properties. Further works are ongoing on the practical use of the strains.

Production, Purification, and Characterization of a Novel Exopolysaccharide from Probiotic Lactobacillus amylovorus: MTCC 8129.

Extracellular polysaccharides (EPS) produced by Lactic Acid Bacteria have an individual effect on the flavour and consistency of novel food materials, as well as potential therapeutic applications. The purpose of this study was to create, improve, and characterise EPS from Lactobacillus amylovorus MTCC 8129. FTIR examination showed the compound's composition (acetyl group, hydroxy group, ring structure) as well as the numerous interlinks between sugar residues, which were then validated by Nuclear Magnetic Resonance Spectroscopy. Thermogravimetric examination showed that the EPS exhibited resistance to heat at a temperature of 640 °C, with antioxidant levels ranging from 70 to 85% and emulsification activity above 50%. Furthermore, it has 180% water holding capacity and 140% oil holding capacity. Based on these findings, it seems that the EPS that was reviewed might potentially be an advantageous addition to the food processing industry.

Production of new functional coconut milk kefir with blueberry extract and microalgae: the comparison of the prebiotic potentials on lactic acid bacteria of kefir grain and biochemical characteristics.

Probiotic foods are recognized for their importance on human health. Kefir is a versatile probiotic food that can be made from non-dairy sources for vegan diet. This study evaluated the addition of microalga Haematococcus pluvialis (0.50% w/v) and blueberry Vaccinium myrtillus (0.50% w/v) extracts to compare their influence on the biochemical properties and the bacterial community of coconut milk kefir through Nanopore-based DNA sequencing. Results revealed that the V. myrtillus increased the microbial diversity in coconut milk kefir with more abundant Proteobacteria species such as Lacticaseibacillus paracasei (22%) and Lactococcus lactis (6.3%). Microalga demonstrated the opposite effect on C, making Firmicutes represent the whole of the microbiota. Biochemical analysis revealed increased fat content in the kefir samples, with the C1 registering 1.62% and the 1.07% in C2, in contrast to the control group's 0.87% fat content. The crude protein content exhibited a decrease in both samples compared to the control group (0.00% and 0.88% versus 1.07%). These findings suggest that fortifying vegan kefir with prebiotics has the potential to induce significant alterations in the kefir microbiota.

Emerging lactic acid bacteria bacteriocins as anti-cancer and anti-tumor agents for human health.

Modern cancer diagnostics and treatment options have greatly improved survival rates; the illness remains a major cause of mortality worldwide. Current treatments for cancer, such as chemotherapy, are not cancer-specific and may cause harm to healthy cells; therefore, it is imperative that new drugs for cancer be developed that are both safe and effective. It has been found that lactic acid bacteria (LAB) have the potential to produce bacteriocins, which could potentially offer a promising alternative for cancer treatment. They have been shown in several studies to be effective against cancer cells while having no effect on healthy cells. More research is needed to fully understand the potential of LAB bacteriocins as anti-cancer medicines, to find the appropriate dose and delivery route, and to conduct clinical trials to evaluate the effectiveness and safety of the products in human patients, as is suggested by this work. Furthermore, LAB bacteriocins may evolve into a significant new class of anti-cancer drugs and food products. Patients with cancer may have a safe and effective alternative treatment option in the form of anti-cancer foods and drugs. Therefore, the aim of this study is to provide an in-depth analysis of the recent breakthroughs and potential future technical advancements of significant bacteriocins that are produced by LAB, how these bacteriocins function, and how these bacteriocins may be utilized as an anti-cancer agent. In addition, the current analysis emphasizes the significant constraints and boundaries that bacteriocins face when they are used as an anti-cancer factor.

Evaluating the Probiotic Profile, Antioxidant Properties, and Safety of Indigenous Lactobacillus spp. Inhabiting Fermented Green Tender Coconut Water.

This study isolated and characterized four indigenous lactic acid bacterial strains from naturally fermented green tender coconut water: Lactiplantibacillus plantarum CWJ3, Lacticaseibacillus casei CWM15, Lacticaseibacillus paracasei CWKu14, and Lacticaseibacillus rhamnosus CWKu-12. Notably, among the isolates, Lact. plantarum CWJ3 showed exceptional acid tolerance, with the highest survival rate of 37.34% at pH 2.0 after 1 h, indicating its higher resistance against acidic gastric conditions. However, all strains exhibited robust resistance to bile salts, phenols, and NaCl, with survival rates exceeding 80% at given concentrations. Their optimal growth at 37 °C and survival at 20 °C and 45 °C underscored adaptability to diverse environmental conditions. Additionally, all strains showed sustainable survival rates in artificial saliva and simulated gastrointestinal juices, with Lact. plantarum CWJ3 exhibiting significantly higher survival rate (70.66%) in simulated gastric juice compared to other strains. Adherence properties were particularly noteworthy, especially in Lact. rhamnosus CWKu-12, which demonstrated the highest hydrophobicity, coaggregation with pathogens and autoaggregation, among the strains. The production of exopolysaccharides, particularly by Lact. plantarum CWJ3, enhanced their potential for gut colonization and biofilm formation. Various in vitro antioxidative assays using spectrophotometric methods revealed the significant activity of Lact. plantarum CWJ3, while antimicrobial testing highlighted its efficacy against selected foodborne pathogens. Safety assessments confirmed the absence of biogenic amine production, hemolytic, DNase, and gelatinase activities, as well as the ability to hydrolase the bile salt. Furthermore, these non-dairy probiotics exhibited characteristics comparable to dairy derived probiotics, demonstrating their potential suitability in developing novel probiotic-rich foods and functional products.

Isolation, genomic analysis and functional characterization of Enterococcus rotai CMTB-CA6, a putative probiotic strain isolated from a medicinal plant Centella asiatica.

Probiotics and their derivatives offer significant health benefits by supporting digestive health, boosting the immune system, and regulating the microbiomes not only of the internal gastrointestinal track but also of the skin. To be effective, probiotics and their derivatives must exhibit robust antimicrobial activity, resilience to adverse conditions, and colonization capabilities in host tissues. As an alternative to animal-derived probiotics, plant-derived lactic acid bacteria (LAB) present promising advantages, including enhanced diversity and tolerance to challenging environments. Our study focuses on exploring the potential of plant-derived LAB, particularly from the medicinal plant Centella asiatica, in improving skin conditions. Through a bacterial isolation procedure from C. asiatica leaves, Enterococcus rotai CMTB-CA6 was identified via 16S rRNA sequencing, whole genome sequencing, and bioinformatic analyses. Based on genomic analysis, antimicrobial-resistance and virulence genes were not detected. Additionally, the potential functions of E. rotai CMTB-CA6 were characterized by its lysates' ability to regulate skin microbes, such as stimulating the growth of Staphylococcus epidermidis while inhibiting that of Cutibacterium acnes, to restore the viability of human dermal fibroblasts under inflammatory conditions, and to demonstrate effective antioxidant activities both in a cell-free system and in human dermal fibroblasts. Our investigation revealed the efficacy of E. rotai CMTB-CA6 lysates in improving skin conditions, suggesting its potential use as a probiotic-derived agent for skin care products. Considering the ecological relationship between plant-inhabited bacteria and their host plants, we suggest that the utilization of E. rotai CMTB-CA6 strain for fermenting its host plant, C. asiatica, could be a novel approach to efficiently enriching bioactive molecules for human health benefits.

Assessment of the effect of lactic acid bacteria fermentation on IgE-/IgG-binding ability and nutritional properties of cow milk.

Cow milk (CM) is an important food source for humans, and food allergy caused by CM has attracted attention worldwide. To our knowledge, systematic studies about the effects of Lactobacillus paracasei, Lactobacillus plantarum, and Pediococcus pentosaceus on the IgE-/IgG-binding ability and nutritional properties of CM are very rare. In this study, L. paracasei, L. plantarum, and P. pentosaceus fermentation on the IgE-/IgG-binding ability was determined by Enzyme-Linked Immunosorbent Assay (ELISA), and the protein quality, amino acid profile, and color were systematically evaluated. The results showed that these LAB strains exhibited higher protein degradation ability, and the IgE reactivity reduction rate was 41.03%-60.00% and the IgG reduction rate was 29.86%-67.20%, respectively. Additionally, the nutritional value was improved obviously, and the color was altered significantly, which was conductive to develop dairy products. These findings provided a theoretical foundation for the development of hypoallergenic dairy products. PRACTICAL APPLICATION: In this study, L. paracasei, L. plantarum and P. pentosaceus could be considered as good potential candidates for solving cow milk allergy owing to their decreased IgE/IgG binding ability andimproved nutritional and sensory properties, which provide a promising strategy to develop hypoallergenic dairy products.

Fortification of yogurt with mulberry leaf extract: Effects on physicochemical, antioxidant, microbiological and sensory properties during 21-days of storage.

Mulberry leaves are medicinal and edible, with many physiological functions. To improve the potential function of yogurt, the effects of mulberry leaf extract (MLE) incorporation on the fermentation kinetics, physicochemical, antioxidant properties, and sensory parameters of yogurt were evaluated. The results showed that 0.1-0.3 % MLE improved the acidification rate and shortened the fermentation process. The addition of MLE significantly increased the values of total titratable acids, water holding capacity (WHC), total phenolic content and antioxidant capacities of the yogurt (p＜0.05). Specifically, the WHC values of 0.1 % MLE added yogurt were 1.33-1.41 times that of the control over 21 days of storage. In addition, MLE changed the texture and sensory quality of yogurt, resulting in light green, more stable products. Compared to the control, the yogurt with an appropriate concentration of MLE (0.1 % and 0.2 %) showed stable microbiological properties, and the survival of lactic acid bacteria in the yogurt was able to maintain a stable probiotic count of 108 CFU/g over 21 days of shelf life. The yogurt containing 0.1 % MLE achieved a good balance between the physicochemical and sensory qualities of the yogurt, and the use of MLE as an ingredient in yogurt production was a step towards the development of healthier dairy products.

Organic acid type in kimchi is a key factor for determining kimchi starters for kimchi fermentation control.

This study was conducted to confirm the effectiveness of kimchi starters (KSs) by investigating their growth characteristics. First, we assessed the growth characteristics of five lactic acid bacteria (LAB) strains (Lactococcus lactis WiKim0124; Companilactobacillus allii WiKim39; and Leuconostoc mesenteroides WiKim0121, WiKim33, and WiKim32) and assessed the effects of different parameters, including organic acids, salinity, acidity, and temperature, on the growth of these LAB. The findings showed that organic acids, particularly acetic and lactic acids that accumulated with the progress in fermentation, were the major players determining the microbial composition of kimchi and the growth of the KSs. Leuconostoc mesenteroides grew well in the presence of acetic and lactic acids than other starts, so it is confirmed that Leuconostoc mesenteroides can dominant in kimchi. In addition, malic acid, which is derived from kimchi ingredients, is used to induce malolactic fermentation by Lactobacillus species, and the progression of malolactic fermentation can be controlled through KSs. Our results suggest that KSs promote the production of organic acids, and the profiling of organic acids, as well as the progress of malolactic fermentation, can be controlled by selecting the suitable KS. Overall, this study demonstrates that kimchi fermentation can be controlled more effectively if the characteristics of KS are understood and used appropriately.

Lactic acid fermentation improves rehydration and emulsifying properties of spray-dried egg yolk powder.

In this study, we investigated the protein structures, powder characteristics, as well as rehydration and emulsifying properties of spray-dried egg yolk powder after short-time lactic acid fermentation (3.5 h). Results indicate that fermentation improved the rehydration and emulsifying properties of yolk powder. Limosilactobacillus reuteri-fermented yolk powder exhibited better wettability due to the porous structure of particles and higher hydrophilicity. Lacticaseibacillus rhamnosus-fermented yolk powder had an enhanced coefficient of stability due to its smaller particles and higher surface charge. The higher water solubility of fermented yolk powder samples is mainly attributed to their lower hydrophobicity and higher zeta potential. The enhanced emulsifying activity of fermented yolk powder samples is primarily related to their increased ß-turn structure and better solubility. Furthermore, fermentation treatment altered powder moisture content and bulk densities, while not affecting its flow behavior and thermal stability. This study provides an effective approach to improving the quality of yolk powder.

Post-acidification of fermented milk and its molecular regulatory mechanism.

The fermented milk products with lactic acid bacteria (LAB) are widely accepted by consumers. During the chilled-chain transportation and storage, LAB in the product keep producing lactic acid, and this will lead to post-acidification, which can affect the flavor, consumer acceptance and even shelf-life of the product. LAB is the determining factor affecting post-acidification. The acid production pathway in LAB and methods inhibiting post-acidification received widespread attention. This review will focus on the post-acidification from the perspective of fermentation starters, including acid production pathway in LAB, main factors and key enzymes affecting post-acidification. Lactobacillus delbrueckii subsp. bulgaricus is a key bacterial species responsible for post acidification in the fermented milk products. The different species and strains presented various differences in process like acid production, acid resistance and post-acidification. Furthermore, multiple factors, such as milk composition, fermentation temperature, and homogenization, also can influence post-acidification. Lactose transport and utilization pathways, as well as its subsequent products metabolic pathway directly influence the post-acidification. F0F1-ATPase, ß-galactosidase, and lactate dehydrogenase are recognized as important enzymes related to post-acidification. The degree of post-acidification is mainly related to the acid production and acid resistance abilities of the fermentation starters, so the key enzymes related to post-acidification are mostly taking part in these two capacities. Recently, some new post-acidification related biomarker genes were found, providing a reference adjusting post-acidification without affecting fermentation rate and bacteria viability. To clarify the post-acidification mechanism at the molecular level will help control post- acidification.

High-Throughput Sequencing Analysis of Microbiota and Enzyme Activities in Xiaoqu from Seven Provinces in Southern China.

Xiaoqu, a pivotal starter in baijiu fermentation, provides the most microflora and enzymes to initiate and maintain baijiu brewing. This study aims to explore the differences in microbiota and enzyme activities among Xiaoqu samples from seven provinces in southern China using high-throughput sequencing, plate isolation, and activity detection. The analyses revealed significant differences in bacterial and fungal communities across the samples. A total of 22 bacterial species and 17 target fungal species were isolated and identified. Predominant bacteria included Bacillus (Bacillus subtilis) and lactic acid bacteria (LABs), while the fungal communities were primarily composed of yeasts (Saccharomyces cerevisiae) and various molds. The activities of α-amylase and glucoamylase varied significantly among the samples, and samples from HN1 and GZ2 exhibited the highest activities. Correlation analyses highlighted the pivotal role of LABs in maintaining acidity and the importance of molds and yeasts in the saccharification and fermentation processes. These findings shed light on the microbial composition and diversity of Xiaoqu and the critical role of microbes in baijiu production. Moreover, they suggested potential microbial resources for developing artificial Xiaoqu via synthetic microbial community in the future, enhancing baijiu fermentation efficiency and overall product quality.

Human breast milk isolated lactic acid bacteria: antimicrobial and immunomodulatory activity on the Galleria mellonella burn wound model.

Introduction: Managing burn injuries is a challenge in healthcare. Due to the alarming increase in antibiotic resistance, new prophylactic and therapeutic strategies are being sought. This study aimed to evaluate the potential of live Lactic Acid Bacteria for managing burn infections, using Galleria mellonella larvae as an alternative preclinical animal model and comparing the outcomes with a common antibiotic. Methods: The antimicrobial activity of LAB isolated from human breast milk was assessed in vitro against Pseudomonas aeruginosa ATCC 27853. Additionally, the immunomodulatory effects of LAB were evaluated in vivo using the G. mellonella burn wound infection model. Results and discussion: In vitro results demonstrated the antimicrobial activity of Lactic Acid Bacteria against P. aeruginosa. In vivo results show that their prophylactic treatment improves, statistically significant, larval survival and modulates the expression of immunity-related genes, Gallerimycin and Relish/NF-κB, strain-dependently. These findings lay the foundation and suggest a promising alternative for burn wound prevention and management, reducing the risk of antibiotic resistance, enhancing immune modulation, and validating the potential G. mellonella as a skin burn wound model.

Continental-scale insights into the sugarbeet diffusion juice microbiomes.

Bacterial contamination of raw diffusion juice poses unique challenges during the sugar extraction process. This study profiled bacterial communities by using full-length 16S rRNA amplicon sequencing and quantified the carbohydrate concentrations in raw diffusion juice samples received from sugar factory regions across the USA and Canada. Juice samples were collected at four time points during the 2021 and 2022 processing campaigns. Firmicutes was the dominant phylum from the raw diffusion juice samples collected during both campaigns and comprised 85.5% of total bacterial abundance. Lactic acid bacteria such as Leuconostoc and Lactobacillus were among the core genera which also dominated the bacterial community in raw diffusion juice. Positive correlations in the abundance of functionally and taxonomically related bacterial communities were identified. During the 2021 campaign, 44 bacterial genera were differentially abundant in raw diffusion juice extracted from sugarbeet roots in Periods 1 to 4. This number declined sixfold during the 2022 campaign to three genera. The concentration of raffinose in raw diffusion juice positively correlated to the relative abundance of Leuconostoc. Furthermore, an in vitro assay was performed to assess the growth dynamics of Leuconostoc mesenteroides in sucrose or raffinose-rich medium and observed the rapid consumption of both carbohydrates by this bacterium. This finding is important for deciphering microbial growth dynamics in raw diffusion juice that can be useful in minimizing sugar loss during the factory processing.IMPORTANCEFindings additionally provide baseline information that can be used to develop mitigation strategies that reduce losses due to microbial contamination of sucrose processing streams.

Antitumor properties of traditional lactic acid bacteria: Short-chain fatty acid production and interleukin 12 induction.

This paper presents an in vitro evaluation of antitumor properties through producing short-chain fatty acids and inducing interleukin 12. In addition, it offers the most important and functional probiotic properties of 24 Lactobacillus gasseri, Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Limosilactobacillus fermentum strains isolated from humans, foods, and fermented foods. To this end, survival in an acidic environment (pH = 2.5), tolerance in bile salt, viability in the presence of pepsin-pancreatin, adhesion percentage, antibiotic resistance, auto-aggregation, and potential percentage of co-aggregation are studied in contact with three human intestinal pathogens. These pathogens are Escherichia coli O157: H7 NCTC 12900, Salmonella enterica subsp. enterica ATCC 13076, and Listeria monocytogenes ATTC 7644. Also, in vitro induction amount of IL-12 in mouse splenocytes is investigated to evaluate antitumor properties by 19 strains of L. gasseri and L. plantarum along with the development of short-chain fatty acids (SCFA) by 5 strains of L. fermentum and L. acidophilus. Gas Chromatography Flame Ionization Detector (GC-FID) and enzyme-linked immunosorbent assay (ELISA) were used to measure short-chain fatty acids and IL-12, respectively. All strains had high viability under acidic conditions. The highest levels of pancreatin and pepsin resistance were found in strains LF56, LF57, LF55, OF, and F and strains LF56, LF57, and A7, respectively. All strains except LF56 had high resistance to bile salts. L. gasseri 54C had the highest average adhesion score (hydrophobicity) of 62.9 % among 19 strains. Despite the susceptibility of different strains of L. plantarum to the tested antibiotics, M8 and M11, S2G, A7, LF55, LF57, and 5G were resistant to kanamycin and chloramphenicol, respectively. Also, 21G was resistant to ampicillin, LF56 to tetracycline and M8, and M11, LF56, and 21G to Erythromycin. In addition, L. gasseri showed moderate resistance to ampicillin, erythromycin, and tetracycline, while L. fermentum ATCC 9338 showed good resistance to ampicillin, erythromycin, and chloramphenicol. In this respect, L. plantarum LF56 and gasseri 54C had the highest average auto-aggregation and co-aggregation against three pathogenic bacteria, respectively. The highest and lowest levels of acetic acid as short-chain fatty acids were produced by L. fermentum 19SH isolated from Horre 41.62 and L. fermentum 21SH from fermented seeds 27.047, respectively. Moreover, L. fermentum, with the OF code of traditional-fermented food origin, produced the most isobutyric acid, butyric acid, and valeric acid, with values of 0.6828, 0.74165, and 0.49915 mmol, respectively. L. fermentum isolated from the human origin with code F produced the most isovaleric acid of 1.1874 mmol. All the tested strains produced good propionic acid except L. fermentum 21SH from fermented seeds. Among strains, L. plantarum M11 isolated from milk and L. gasseri 52B from humans had the highest in vitro induction of IL-12, which is probably related to their cell wall compositions and structure.